Objectives: Mesenchymal stem cells (MSCs) isolated from various tissues have been used for clinical application in tissue engineering and regenerative medicine. To date, the most common source of MSCs has been bone marrow. However, the bone marrow aspirate is an invasive and painful procedure for the donor. Thus, the identification and characterization of alternative sources of MSCs are of great importance.
Aim of this study was isolation and characterization of MSCs from human dental pulp (DPSC).
Materials and methods: The Ethical Committee of Medical Research Office Hamad Medical Corporation, Doha, Qatar, approved our experimental protocol. Human dental pulp was extracted from premolars and third molars ( n = 19) of healthy subjects 10–21 years of age after informed consent. The pulp was digested in a solution of 3 mg/mL collagenase type I and 4 mg/mL dispase for 1 h at 37 °C. After filtration using 70-mm cell strainers (Falcon), cells were cultured in Dulbecco’s Modified Eagle Medium containing 20% mesenchymal cell growth supplement and antibiotics (100 U/mL penicillin, 100 mg/mL streptomycin, and 0.25 mg/mL amphotericin B) at 37 °C under 5% CO 2 . Cultures were treated with osteoinductive medium for characterization and differentiation. The medium changed twice weekly.
Results: The stem cells were detected by electronic microscope by visualization after period between 2 and 3 weeks. The primary colonies were grown for next 2 weeks and then when reached confluence by positive Alizarin Red Staining reaction they showed their osteogenic differentiation capability. Also a reaction showed strong mineralization of new formed tissue in vitro.
Conclusions: In this study we showed that DPSC constitute an ideal source of osteoblasts and mineralized tissue for bone regeneration. The main advantage of using DPSC is non-morbidity access and can be obtained noninvasively from deciduous teeth that are routinely extracted and generally discarded as medical waste without any ethical concerns.
Conflict of interest: None declared.